Background: Osteoarthritis (OA) characterized by progressive cartilage degeneration and chronic pain is hindered by the vicious inflammation-pain cycle. Nonsteroidal anti-inflammatory drugs (NSAIDs) can only alleviate clinical symptoms. Although electrical stimulation of the vagus nerve has achieved some results, the toxicity of the electrodes and the secondary damage caused by dismantling limited its clinical application. Methods: A "sono-piezoelectric-bioelectricity-neuroimmune" cascade modulation strategy based on ultrasound-driven piezoelectric ZnO nanoparticles was established to attenuate osteoarthritic neurogenic inflammation and pain with inhibited cartilage degradation. Results: The "sensory neuron-cholinergic anti-inflammatory pathway" reflex arc was activated by the dynamically and spatially-temporally programmed sono-piezoelectric field (0.7 V/36 μA peak) within deep joint tissues by targeting α7nAChR-P2RX7 neuroimmune axis. Consequently, our sono-piezoelectric neuroimmune modulation strategy significantly up-regulated α7nAChR expression, and synchronously inhibited pain mediator CX3CL1, opposed macrophage infiltration, inhibited P2RX7-mediated IL-1β/IL-6 inflammatory storm, restored IL-1β-injured chondrocyte activity and migration capacity, activated stromal genes (Col2a1) for matrix synthesis, and inhibited cartilage degradation-related MMP13 expression. All these actions re-established the balance of glycosaminoglycan (GAG)/deoxyribonucleic acid (DNA) metabolism to remodel joint immune homeostasis, and activated the cholinergic pathway to break the vicious cycle of "inflammation-pain", which restored mechanical pain threshold (Von Frey) and weight-bearing capacity to near-normal levels and reconstructed tidal structures in a rat OA model. Conclusions: Our pioneering "sono-piezoelectrical signal-neural reflex-immunomodulation" cascade strategy for regulating neuroinflammatory reflex-arc-mediated α7nAChR-P2RX7 axis provide deep insights into OA-represented neuroinflammatory diseases.

Hyperuricemia is closely related to gout, renal disease and cardiovascular disease, and the development of high-performance tools for serodiagnosis and drug efficacy assessment is of great practical significance. In this study, a dual-channel hydrogel film (CHI-PMoV/VI-UOx) is developed for dual-channel detection of uric acid and showed a good quantification performance and linearity in the physiological uric acid level. Combined with the self-developed Android application and multi-channel strategy, it shows high serodiagnostic accuracy of 96.0% and great potential in drug screening.

ObjectiveThis cross-sectional study aimed to examine the association between combined healthy lifestyle factors and the prevalence of osteoarthritis (OA) among middle-aged and older U.S. adults using nationally representative data from the National Health and Nutrition Examination Survey (NHANES).MethodsData were collected from 15,617 adults participating in NHANES (2005-2020). A healthy lifestyle score (range 0-6) was constructed based on six modifiable factors: smoking status, alcohol consumption, physical activity, diet quality, waist circumference, and sleep duration. Weighted multivariable logistic regression models were utilized to analyze the relationships between healthy lifestyle scores and the prevalence of OA, controlling for demographic, socioeconomic, and health-related variables. Robustness was assessed through stratified, interaction, and sensitivity analyses, including propensity score adjustment, E-value analysis, and restricted cubic spline modeling.ResultsAmong 15,617 participants, OA was present in 2,275 individuals. In the crude model, the association between the healthy lifestyle score and OA was not statistically significant for the per-factor trend. After adjustment for covariates, a higher healthy lifestyle score was significantly associated with lower odds of OA. Participants with 5-6 healthy lifestyle factors had 31% lower odds of OA, and each additional factor was associated with 7% lower odds of OA. Key factors associated with lower odds of OA included non-smoking, optimal waist circumference, and adequate sleep. The association was more pronounced in adults under 60 years. Sensitivity analyses confirmed a consistent inverse association between the lifestyle score and OA odds.ConclusionsOur findings suggest that adherence to a greater number of healthy lifestyle factors may be associated with lower odds of OA, particularly among younger adults. Notably, smoking avoidance, maintaining optimal waist circumference, and adequate sleep duration demonstrated particularly robust inverse associations. These results highlight the potential of integrated lifestyle modifications for OA-related public health strategies at both individual and population levels.

Bone metastasis is a major complication of breast cancer, characterized by osteolytic destruction mediated by excessive osteoclast activation. Current anti-resorptive therapies primarily target osteoclasts but have limited impact on the tumor-bone microenvironment vicious cycle that drives bone destruction. This study evaluated the therapeutic efficacy of a novel pan-B-cell lymphoma 2 inhibitor, Sabutoclax, in a breast cancer-induced model osteolysis and explored potential mechanisms associated with its effects. At the cellular level, we assessed the effects of Sabutoclax on the proliferation, invasion, migration, and apoptosis of cancer cells. We observed that Sabutoclax treatment was associated with inhibition of RANKL-induced osteoclast differentiation, RANKL-induced acid secreactivated protein kinase/extracellular signal-regulated kinase signaling, as well as reduced nuclear translocation and expression of nuclear factor of activated T-cells cytoplasmic 1 (NFATc1). In the animal experiment, an orthotopic breast cancer osteolysis model in the tibia of nude mice was established. The in vivo efficacy of Sabutoclax was evaluated. This study found that Sabutoclax effectively prevents breast cancer-induced osteolysis, which may involve a dual mechanism, suppressing breast cancer cell functions and targeting osteoclast differentiation and acid secretion. And the present study only shows that Sabutoclax is associated with ROS reduction, mitochondrial perturbation, and suppression of ERK/NFATc1 signaling. Sabutoclax treatment was associated with both decreased protein expression and reduced nuclear translocation of NFATc1. Future studies could focus on comprehensive evaluation of its pharmacokinetic properties, systemic toxicity, and therapeutic efficacy in more clinically relevant metastatic models to establish its potential application in breast cancer-induced osteolytic bone destruction.

AimOsteoarthritis (OA) is increasingly understood as a condition influenced by the diversity of chondrocytes and immune system dysfunction; however, the specific molecular elements connecting distinct chondrocyte populations to the immune environment remain inadequately explored. The protein CORO1A, associated with the cytoskeleton and implicated in immune regulation, has not been thoroughly investigated in the context of OA or through single-cell analysis.ApproachWe employed multi-omics techniques alongside single-cell RNA sequencing, supported by experimental validation using human OA cartilage samples, a murine model of medial meniscus destabilization (DMM), and in vitro inflammatory chondrocyte models to elucidate the expression patterns and functional significance of CORO1A in OA.FindingsThe CORO1A gene is recognized as significantly expressed in osteoarthritic cartilage. Transcriptomic evaluations reveal that CORO1A is predominantly expressed in inflammatory and proliferative chondrocyte subpopulations, while showing lower levels in other chondrocyte types, underscoring the specificity of these groups. The upregulation of CORO1A is associated with increased infiltration of various immune cells, including M2 macrophages, plasma cells, and natural killer cells, and is linked to the activation of IL-6/JAK-STAT3 and TNF-α/NF-κB signaling pathways. Additionally, experimental data indicate that CORO1A is markedly elevated in the cartilage regions affected by osteoarthritis in both humans and mice. In terms of its function, the expression of CORO1A in primary chondrocytes is enhanced by inflammatory stimuli, while its suppression leads to reduced MMP13 expression and diminished chondrocyte migration. Conclusion: This study identifies CORO1A as a regulator specific to subpopulations within the immune microenvironment, connecting immune remodeling to both inflammatory and proliferative chondrocytes in OA. Considering the diversity of chondrocytes, the presence of immune cells, and catabolic processes, our results emphasize the significance of CORO1A as a mechanistic factor contributing to cartilage degradation, suggesting its potential as a therapeutic target for OA.

The clinical management of osteosarcoma faces critical challenges, including postoperative recurrence and metastasis of residual tumor cells, chemotherapy resistance, and impaired self-repair capacity due to extensive bone defects following tumor resection. Herein, we develop a novel functionalized 3D-printed scaffold with NIR-II responsiveness (BGS/I-LDH@MgO2), designed to simultaneously address the dual needs of inhibiting osteosarcoma recurrence and promoting bone regeneration. This scaffold consists of 3D-printed bioactive glass scaffold (BGS) and MgO2-modified ZnAl-layered double hydroxides (ZnAl-LDHs) intercalated with 5-iodo-isophthalic acid (I-IPA). Under NIR-II irradiation, the scaffold effectively triggers a photodynamic therapy (PDT) effect to eliminate osteosarcoma cells. Notably, the incorporation of MgO2 enables oxygen release within the tumor microenvironment, alleviating hypoxia and enhancing PDT efficacy for superior antitumor performance. Furthermore, the degradation of ZnAl-LDHs and MgO2 releases Mg2+ and Zn2+ ions and generates a mildly alkaline microenvironment, which collectively facilitate the osteogenic differentiation of bone marrow mesenchymal stem cells and accelerate the process of bone healing. This functionalized 3D-printed scaffold demonstrates excellent anti-tumor and osteogenic properties, showing great promise for the treatment of osteosarcoma-associated bone defects.

Background Osteosarcoma (OS) is the most prevalent primary cancer of the bone. Metastasis and chemoresistance are the major obstacles to the improvement of OS prognosis, in which N-6-methyladenosine (m(6)A) modification plays an important role, but the exact molecular mechanisms are still unclear. Methods MeRIP-seq and RNA-seq were conducted on OS and paired adjacent normal tissue samples, which determined CACNA1E as a key m(6)A-modified molecule. In vitro and in vivo models were established to evaluate the function of CACNA1E on OS growth, metastasis, and methotrexate (MTX) resistance, and to explore the upstream regulators and downstream effectors of CACNA1E. Results CACNA1E exhibited notable m(6)A hypermethylation and upregulated expression in OS than adjacent normal tissues. CACNA1E knockdown effectively hindered OS growth, lung metastasis, and MTX resistance. METTL3, an m(6)A "writer" boosted the mRNA stability of CACNA1E through m(6)A modification, and this process was recognized and enhanced by IGF2BP2, an m(6)A "reader". WNT7B was identified as a downstream molecule of CACNA1E. CACNA1E facilitated OS progression and MTX resistance by enhancing the non-canonical Wnt/Ca2+ signaling through transcriptionally activating WNT7B. Furthermore, a novel combination treatment of targeted inhibition of CACNA1E with MTX had a synergistic effect on suppressing OS progression. Conclusions Collectively, our findings uncover that METTL3-mediated m(6)A modification of CACNA1E contributes to OS progression and chemoresistance through enhancing WNT7B-mediated non-canonical Wnt/Ca2+ signaling. Targeted inhibition of CACNA1E in combination with MTX may be a promising alternative therapeutic strategy for patients with MTX-resistant OS.

Objective Gout and hyperuricemia (HUA) are common metabolic disorders associated with increased all-cause and cardiovascular disease (CVD) mortality, and gut microbiota-modulating diets may influence prognosis. We aimed to evaluate whether the Dietary Index for Gut Microbiota (DI-GM) is associated with all-cause and CVD mortality among adults with gout or HUA. Methods In this prospective cohort study, we analyzed data from 5,325 adults with gout or hyperuricemia enrolled in the 2007–2018 National Health and Nutrition Examination Survey (NHANES). DI-GM scores were calculated using 24-hour dietary recall data, incorporating 14 predefined food components (10 beneficial and 4 harmful to gut microbiota). Mortality status was ascertained through linkage to the National Death Index up to December 31, 2019. Cox proportional hazards models were applied to estimate hazard ratios and 95% confidence intervals, adjusting for demographic, lifestyle, and clinical factors. Sensitivity analyses, interaction tests, and stratified analyses were conducted to assess robustness. Results During a median follow-up of 6.5 years (35,625 person-years), 603 deaths occurred, including 197 from CVD. Higher DI-GM scores were associated with significantly lower mortality risk. Participants with DI-GM scores ≥6 had a 33% reduced risk of all-cause mortality and a 45% reduced risk of CVD mortality compared to those with scores ≤ 4. Each one-point increase in DI-GM score was associated with an 8% decrease in all-cause mortality and a 13% decrease in CVD mortality. Results were consistent across all sensitivity analyses. Conclusions In individuals with gout or hyperuricemia, higher DI-GM scores were independently associated with reduced all-cause and CVD mortality. These findings suggest that gut microbiota-friendly dietary patterns may provide prognostic value and offer a potential target for dietary interventions in this high-risk population.

ObjectiveThe purpose of this study was to reveal the genetic correlation of RANKL polymorphisms with bone metastasis in breast cancer patients.MethodsIn this study, 139 bone metastasis patients and 152 no metastasis were included as the case and control groups. Real-time polymerase chain reaction (PCR) and allelic discrimination method were respectively applied for the genotyping of rs7325635 and rs2277438. Polymorphism genotype and allele frequencies were compared by χ2 test between the two groups. The risk of bone metastasis development caused by RANKL genetic variants was evaluated by odds ratio (OR) with 95% confidence interval (95%CI). The linkage of two polymorphisms was examined by Haploview. Binary and multivariate logistic analyses were used to optimize the results.ResultsRs2277438 GG genotype and G allele frequencies were significantly higher in bone metastasis patients than that in no-metastasis patients (P < 0.05); they were significantly correlated with the increased risk of bone metastasis occurrence (GG: OR = 2.065, 95%CI=1.104-3.863; G: OR = 1.486, 95%CI=1.068-2.068). Compared with the G–G haplotype, the A–A haplotype was found to significantly reduce the risk of bone metastasis in breast cancer patients (OR = 0.647, 95%CI=0.436-0.959). The multivariate logistic analysis indicated that family history, Ki67, and rs2277438 were positively correlated with bone metastasis, but menopausal state, clinical staging, and rs7325635 were negatively correlated with bone metastasis in breast cancer.ConclusionThe RANKL rs2277438 variant may be a potential genetic susceptibility factor associated with bone metastasis risk in breast cancer patients, though further functional validation is warranted. Rs7325635 was not independently associated with bone metastasis. The linkage disequilibrium between these two polymorphisms and their combined haplotype effect may play a role in bone metastasis susceptibility.

Objective To comprehensively and systematically elucidate the role of the regulatory network of gutassociated microbiota metabolites (GMMs) on the onset of rheumatoid arthritis (RA) while identifying important molecular targets and choosing potential drug molecules. Methods The differentially expressed genes derived from the RA gene chip data were combined with the genes from the weighted gene co-expression network analysis module, utilizing databases such as SwissTargetPrediction (STP), Similarity Ensemble Approach (SEA), and GutMgene to derive the target gene set associated with gut microbiota metabolites. After that, the intersection between the above data with RA-associated genes obtained from GeneCards, OMIM and CTD was identified. GO/KEGG functional annotation and topological evaluation of protein-protein interaction networks were used to filter key targets. A multi-dimensional regulatory framework of "micro-organism-metabolite-gene target" was established, and drug characteristics and toxicity profiles were assessed using SwissADME and ADMETlab platforms. Molecular docking studies were carried out to validate the interaction of the selected lead compounds and core targets. At the same time, using a RA cell model, the in vitro activity of the candidate drugs was evaluated in synovial fibroblasts through half-maximal inhibitory concentration (IC50), cell counting kit-8(CCK-8) proliferation assay, clonogenic assay and other methods. Results A total of 40 common genes were successfully identified through multi-dimensional analysis, in which IL6, TNF, IL1β, AKT1 and TP53 were confirmed to be key regulatory factors. The pathway enrichment results indicated that these genes were significantly involved in everal critical pathological processes of RA, including PI3K-Akt, MAPK, TNF signaling transduction, Th17 cell differentiation and NOD-like receptor pathways. An interaction network containing 154 bacterial metabolites, 1, 518 metabolism-related targets and 1, 933 disease-related targets was constructed, which highlighted the regulatory role of Flavonifractor plautii and Blautia sp. in modulating core targets. These microbiota regulated the action mode of the core target via tryptophan and tyrosine metabolic pathways. The assessment of efficacy and toxicity led to the identification of lead compounds with good drugability, which exhibited a significant binding activity to targets like IL6 and TNF. This compound had an IC50 value of 40.85 μmol/L for synovial fibroblasts based on in vitro studies and could inhibit cell proliferation and colony formation in a dose-dependent manner. When the concentration was no more than 80 μmol/L, the survival rate of cells was over 80%, showing excellent safety characteristics. Conclusion This study systematically elucidates the key mechanism by which the gut microbiota regulates the immune-inflammatory signaling axis in the pathogenesis of RA through its metabolites. The identified core gene network and the candidate drug 3-(4-Hydroxyphenyl) propionic acid provide a scientific basis for precision treatment strategies of RA and open up new avenues for drug repurposing research.

Non-traumatic osteonecrosis of the femoral head (NONFH) is a disabling disease mainly caused by glucocorticoid use and alcohol abuse, characterized by local impairment of blood supply and limited bone repair; however, the composition and regulatory mechanisms of specialized endothelial cells (ECs) in its microenvironment remain unclear. In this study, we used single-cell RNA sequencing (scRNA-seq) to identify four EC subsets, including type L, type H, type R, and arterial ECs. Type H and type R ECs were primarily enriched in cell adhesion and angiogenesis pathways and represented key subsets that promote bone repair in NONFH. Their abundance increased during the mid-stage of NONFH but declined in the late stage. Stromal cells in the femoral head interacted with type H and type R ECs through multiple ligand–receptor axes, maintaining their function. Further screening identified PDK4 as a critical regulator of type H and type R EC abundance. PDK4 not only promotes endothelial fatty acid oxidation but also participates in iron homeostasis and angiogenesis. In vitro experiments showed that PDK4 knockdown significantly impaired the angiogenic phenotype of ECs, which was associated with suppressed fatty acid oxidation, highlighting PDK4’s role in sustaining the activity and function of type H and type R ECs under disease conditions. Peripheral blood transcriptome analysis revealed that the dynamic expression pattern of PDK4 across NONFH stages was consistent with the single-cell results, suggesting potential value for stage-specific diagnosis. By integrating single-cell and peripheral blood transcriptome analyses with in vitro experiments and metabolomic validation, this study elucidates the dynamic changes of specialized ECs and the PDK4-mediated metabolic regulatory mechanisms during NONFH progression. These findings provide new insights into vascular repair and potential therapeutic strategies for NONFH.

Objective Diabetic osteoporosis is a secondary complication of diabetes mellitus, characterized by reduced bone mass, increased bone fragility, and impaired fracture healing. However, the mechanisms underlying diabetic bone loss remain to be fully elucidated. More importantly, there is an urgent need to identify therapeutic agents that not only lower blood glucose levels but also alleviate bone loss. Therefore, this study aims to investigate the mechanisms of diabetes-associated bone loss and to explore potential therapeutic agents. Methods We established a mouse model of type 2 diabetes (T2D) induced by streptozotocin and a high-fat diet (HFD). Bone mass and osteoclast numbers were assessed using micro-CT and TRAP staining. In vitro, the effects of MSDC-0160 (MSD) on osteoclast differentiation and function were evaluated through TRAP staining and bone resorption assays. To elucidate the molecular mechanisms underlying MSD-mediated inhibition of osteoclastogenesis, qPCR, Western blotting, and immunofluorescence staining were performed. Finally, micro-CT scanning and immunohistochemical staining were conducted to examine the effects of MSD on bone microstructure and the bone microenvironment in T2D mice, as well as to clarify specific mechanism of action. Results T2D mice exhibited significant bone loss and enhanced osteoclast activation. Moreover, mitochondrial pyruvate carrier (MPC) activity was elevated in osteoclasts of T2D mice. Given the potential for mitigating diabetic bone loss by inhibiting MPC activity, we selected MSD, a novel insulin sensitizer that also serves as an MPC inhibitor. Further detailed investigations revealed that MSD suppresses osteoclast differentiation and function by reducing the energy supply required for osteoclast maturation. This effect results from impaired mitochondrial oxidative phosphorylation (OXPHOS) and reduced mitochondrial biogenesis. In vivo administration of MSD significantly ameliorated bone loss and reduced osteoclast numbers in T2D mice. Conclusion Our findings indicate that the bone loss in T2D mice is associated with excessive osteoclast activation, where MPC playing a crucial role in osteoclast differentiation and maturation. MSD, a novel insulin sensitizer, mitigates diabetic bone loss by suppressing MPC activity in osteoclasts.

Ferroptosis, driven by redox imbalance, plays a critical role in osteoarthritis (OA) progression. Although antioxidant nanozymes hold therapeutic potential, designing highly efficient and targeted systems to inhibit ferroptosis remains challenging. Here, we developed a 2D nitrogen-doped graphene-like nanomesh (NGM) loaded with asymmetric and highly exposed Fe single atoms, carried with the cartilage-targeting WYRGRL peptide and siRNA (siMMP13) to form Fe SAzymes (si-FeSA/NGM-W) as ferroptosis inhibitors to alleviate OA. By mixed molten salt and Zn removal, exfoliating Zn-ZIF into an ultrathin 2D hierarchical porous NGM with topological defects and hierarchical structure, we created a scaffold for anchoring asymmetric and highly exposed Fe single atoms. The abundant Fe-N4-Cl coordination active sites then introduce strain and defects, which facilitate electron transfer, enhance radical adsorption, and lower reaction barriers, thereby augmenting multi-enzyme (SOD/CAT/GPx) activities. This enables the functionalized si-FeSA/NGM-W to target cartilage, where it inhibits ferroptosis by downregulating MMP13, upregulating GPX4, restoring mitochondrial function, and modulating inflammation, ultimately achieving targeted OA therapy. Mechanistically, this process involves suppression of the IL-17 pathway and enhancement of glutathione metabolism. This work presents a targeted nanozyme platform for precise OA therapy via ferroptosis inhibition.

ObjectiveThis study aims to elucidate the molecular mechanisms through which PET microplastics (PET-MP) influence osteoarthritis (OA) pathogenesis by integrating network toxicology, machine learning, and in vitro experimental validation.MethodsDifferential gene expression analysis and WGCNA were applied to multiple OA datasets to identify disease-related targets. PET-MP biological targets were predicted via ChEMBL, SwissTargetPrediction, and PharmMapper. Overlapping targets were screened using machine learning algorithms, and molecular docking was performed to assess binding interactions. In vitro validation including immunofluorescence, qRT-PCR, and Western blot was conducted in PET-MP-treated chondrocytes.ResultsA total of 452 PET-associated targets were identified, with 12 core PET-MP-OA genes established through intersection analysis. Functional enrichment implicated the NF-κB and IL-17 signaling pathways. Machine learning screening based on feature importance and SHAP values prioritized six hub genes: AKR1A1, INSR, KIF11, MMP1, KCNN4, and TK1. Molecular docking generated predicted AutoDock Vina scores ranging from −3.893 to −7.434 kcal/mol. In vitro experiments validated upregulation of AKR1A1, MMP1, KCNN4, KIF11, and TK1, and downregulation of INSR in chondrocytes, consistent with bioinformatics predictions.ConclusionPET-MP may promote OA progression by disrupting molecular pathways related to inflammation, oxidative stress, and cartilage degradation. The identified hub genes offer new insights into microplastic toxicology in joint disease and represent potential therapeutic targets and biomarkers for PET-MP-induced OA.

Pyroptosis inhibition via Fe single-atom nanozymes is promising for inflammation therapy, but the common Fe-N4 configuration restricts oxygen intermediate desorption and lacks cooperative sites, thus limiting catalytic performance. To overcome this, we develop a FeMn dual-atom nanozyme supported on oxygen-nitrogen-doped bamboo-like carbon nanotubes (FeMnDA/BCNT). Through the precise alignment of Fe and Mn 3dz2 orbital energy levels by the electron-delocalized BCNT support in the FeMn-N/O active center, thereby lowering the dissociation energy barrier for *O2 or *H2O molecules, promoting O─O bond cleavage to bypass toxic ─OOH species, and thus accelerating the enzyme-like kinetics. Combined with a hierarchical porous bamboo-like structure of the BCNT that enhances high specific surface, atom exposure, and mass transfer, the FeMnDA/BCNT nanozymes exhibit potent superoxide dismutase (SOD), catalase (CAT), and glutathione peroxidase (GPx)-like activities. Further encapsulation with a macrophage membrane ([MM]FeMnDA/BCNT) confers excellent biocompatibility and active targeting ability toward inflammatory sites. The resulting [MM]FeMnDA/BCNT nanozymes target the inflammatory microenvironment, scavenges ROS, restores mitochondrial function, and suppresses NLRP3 inflammasome activation, thereby inhibiting pyroptosis. In vivo, [MM]FeMnDA/BCNT nanozymes show good biocompatibility and efficacy in treating osteoarthritis, acute liver injury, and acute kidney injury. This work provides a novel strategy for inflammatory disease therapy using a biomimetic dual-atom nanozyme.

Background:As a main component of the tissue microenvironment, the extracellular matrix (ECM) provides an instructive niche that regulates stem cell differentiation. Biomimetic ECM collagen hydrogels have been well regarded as ideal scaffold for cartilage tissue engineering due to their ability to promote chondrogenic differentiation of bone marrow mesenchymal stem cells (BMSCs). However, the molecular mechanisms underlying this inductive effect remain incompletely elucidated. Methods:In the present study, mRNA microarray analysis was performed to investigate the molecular mechanisms involved in collagen hydrogel-mediated chondrogenic induction. Real-time quantitative polymerase chain reaction (RT-qPCR), western blot, immunofluorescence analyses, etc. were used to validate the relevant pathways. Results:The results demonstrated that POU class 5 homeobox 1 (POU5F1), a transcription factor associated with stem cell differentiation and autophagy, was highly expressed in cells induced by collagen hydrogel. Differentially expressed genes (DEGs) were predominantly enriched in the AMP-activated protein kinase (AMPK) / mammalian target of rapamycin (mTOR) signaling pathway. Knockdown of POU5F1 suppressed activation of the AMPK/mTOR pathway and subsequently reduced intracellular autophagic flux, leading to impaired cartilage regeneration. These effects were partially reversed by treatment with rapamycin (RaPa), an mTOR inhibitor. Conclusions:The findings highlight a critical role of autophagy in chondrogenic induction mediated by biomimetic ECM collagen hydrogel and provide mechanistic insight that may inform the rational design optimization of cartilage repair biomaterials.

Directional migration and differentiation of stem cells in situ are necessary for cartilage tissue regeneration. To regenerate cartilage tissue similar to natural one, transplanted scaffolds are required to have favorable stiffness, biocompatibility, and the ability to recruit and induce stem cells. Herein, a three-dimensional (3D) bio-printed Gum Karaya Methacrylate (GK-MA) bioink functionalized with EPLQLKM (E7) peptide was developed to form GK-MA-E7 hydrogel for cartilage tissue engineering applications. Among them, GK-MA exhibits desirable mechanical strength and biosafety, while the E7 peptide acts as an anchoring site for the recruitment of bone marrow mesenchymal stem cells (BMSCs). The physicochemical properties of the bioink can be precisely regulated by adjusting the photopolymerization conditions. Compared to Gum karaya (GK) and GK-MA scaffolds, the GK-MA-E7 construct significantly enhances the biological behavior of BMSCs, as reflected in improved adhesion, proliferation, homing efficiency, and chondrogenic commitment. These effects are corroborated by a pronounced increase in glycosaminoglycan (GAG) deposition and the elevated transcription of cartilage-specific markers such as Col2a1, SOX9, Col1a1, and ACAN. Further, the photo-cross-linkable 3D-printed GK-MA-E7 hydrogels embedded with BMSCs led to notable cartilage regeneration in vivo by the end of the 8-week therapeutic regimen, with the score elevated to approximately three times that of the control group. This work proposes an alternative approach for the development of photo-cross-linkable, biodegradable, and injectable scaffolds derived from native polysaccharides, offering the potential for minimally invasive strategies in cartilage tissue regeneration.

Human Papillomavirus (HPV) types 16 and 18 are well-established causative agents in cervical cancer. However, the mechanism of malignant transformation remains unclear. Although epithelial-mesenchymal transition (EMT) is regulated by Ca2+ signaling, the functions of transient receptor potential canonical (TRPC) channel in cervical cancer have not been reported. Herein, employing multiple biological approaches, we first revealed that HPV16 and HPV18 infections significantly upregulated the expression of TRPC3 that orchestrated Ras-MAPK and MEK-ERK pathways in the abnormal transformation of cervical epithelial cells. Our transcriptomic sequencing of HPV-infected cervical epithelial cells with depletion of TRPC3 suggested the significant influence of TRPC3 on genes involved in the process of epithelial-mesenchymal transition (EMT). Consistently, inhibition of TRPC3 successfully suppressed HPV-triggered cell viability and EMT. Moreover, we found TRPC3 maintained the viability of HPV - infected cells by suppressing excessive MAPK activation through regulating Ras GTPase - activating protein 4 (RASA4), which was validated by the detection of phosphorylated ERK1/2 (p-ERK1/2).These findings were further confirmed in the HPV-infected female BALB/c mice, highlighting TRPC3 as a key hub in mediating transformation This study advances the knowledge about the Ca2+ signaling-related molecular mechanism underlying HPV-driven malignant transformation. Targeting TRPC3 may have broader therapeutic implications. This study reveals that the TRPC3 channel is essential for HPV-driven cervical cancer. TRPC3 promotes malignant transformation by regulating the RASA4/MAPK pathway and EMT. Targeting TRPC3 could be a new therapeutic strategy.

Rationale: Bone defects pose a persistent challenge in orthopedic medicine due to their limited self-repair capacity. Although guided bone regeneration scaffolds have shown therapeutic potential, their clinical efficacy remains constrained by their suboptimal osteoinductive capability. Methods: Herein, we developed biodegradable piezoelectric polyhydroxybutyrate-barium titanate (PHB-BT) nanofiber scaffolds capable of generating synergistic piezoelectric stimulation for bone repair when integrated with low-intensity pulsed ultrasound (LIPUS). Results: Compared with conventional PHB scaffolds, ​PHB-BT nanofiber scaffolds​ ​showed enhanced piezoelectric properties​ and ​excellent biocompatibility, ​thereby facilitating​ sustained osteogenic activity. ​In vitro​ studies revealed that these scaffolds ​significantly promoted​ the osteogenic differentiation of bone marrow mesenchymal stem cells under LIPUS stimulation. ​Notably, ​in vivo​ evaluations ​demonstrated​ that these scaffolds ​substantially accelerated bone defect repair​, with complete scaffold degradation observed after eight weeks. Mechanistically, PHB-BT nanofibers improved osteogenesis via activating the Ca2+/calcineurin/nuclear factor of activated T-cells signaling pathway in response to ultrasound stimulation. Conclusions: These findings have significant implications for the design of next-generation, implantable electrical stimulators capable of providing sustained electromechanical cues for personalized bone tissue engineering applications.

BackgroundMyocardial infarction (MI) is a myocardial necrosis event caused by an unstable ischemic state that reduces life expectancy primarily through cardiac functional impairment and cardiomyocyte death. The present study aims to investigate the genetic mechanisms underlying MI by integrating expression quantitative trait loci (eQTLs) and Mendelian randomization (MR) analyses.MethodsWe comprehensively analyzed independent MI datasets from the Gene Expression Omnibus database. The relationships between MI and the differentially expressed genes were evaluated through differential expression, eQTL, and MR analyses. Additionally, GO and KEGG enrichment analyses were performed to clarify the functional pathways of the candidate genes, and gene set enrichment analysis (GSEA) was used to identify the genes associated with MI. An in vitro model of MI was established by subjecting AC16 cells to oxygen and glucose deprivation, and the gene expression levels were validated through reverse transcription quantitative polymerase chain reaction (RT-qPCR).ResultsBy comparing the results from the MR analysis and mRNA expression profiles, we identified 13 overlapping genes: MRPL35, SNUPN, ADM, BCL6, BNIP3L, CMTM2, DGAT2, HSPA6, IER3, IFNGR1, PLAUR, SERPINB8, and VNN2. The GO and KEGG enrichment analyses revealed that these genes participate in essential biological processes, including mitochondrial apoptotic and mitochondrial organization regulatory pathways. GSEA demonstrated that the candidate genes were enriched in the NOD-like signaling pathways; immunological responses; and lysosomal, ribosomal, and metabolic pathways related to MI. Furthermore, the gene expression levels were verified through RT-qPCR.ConclusionThis study highlights the potential of specific molecular pathways for targeted treatment of MI. Our work also warrants additional research efforts to elucidate the genetic mechanisms of MI.